title: Development of a high speed laser scanning confocal microscope with an acquisition rate up to 200 frames per second.
abstract: There has been an increasing interest for observing fast biological phenomena such as cell movements in circulations and action potentials. The laser scanning confocal microscopy offers a good spatial resolution and optical sectioning ability to observe various in vivo animal models. We developed a high speed laser scanning confocal microscope capable of acquiring 512 by 512 pixel images at 200 fps (frames per second). We have incorporated a fast rotating polygonal scanning mirror with 128 facets for the X-axis scanner. In order to increase the throughput of the Y-axis scanner, we applied a bi-directional scanning method for vertical scanning. This made it possible to scan along the Y-axis two times during each scanner motion cycle. For the image acquisition, we used a custom photomultiplier tube amplifier with a broad frequency band. In addition, custom imaging software was written for the new microscope. In order to verify the acquisition speed of the developed confocal microscope, a resolution target moving at a series of constant speeds and a sedated mouse with slight movements due to heartbeats were observed. By comparing successive frames, the frame acquisition speeds were calculated.
authors.forename: S
authors.initials: S
authors.lastname: Choi
authors.forename: P
authors.initials: P
authors.lastname: Kim
authors.forename: R
authors.initials: R
authors.lastname: Boutilier
authors.forename: M Y
authors.initials: MY
authors.lastname: Kim
authors.forename: Y J
authors.initials: YJ
authors.lastname: Lee
authors.forename: H
authors.initials: H
authors.lastname: Lee
mesh_terms: D000818:Animals; D051379:Mice; D018613:Microscopy, Confocal / Q000295:instrumentation*; D012867:Skin / Q000033:anatomy & histology; D013997:Time Factors